建立一种用于检测牛精液中牛α疱疹病毒 1 的液滴数字 PCR 检测方法。
Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen.
机构信息
Department of Veterinary Medicine, College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang, 524088, Guangdong, China.
Technology Center, Hohhot Customs District, Hohhot, 010020, Inner Mongolia, China.
出版信息
BMC Vet Res. 2022 Apr 2;18(1):125. doi: 10.1186/s12917-022-03235-2.
BACKGROUND
Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is one of the most important contagious diseases in bovine. This is one of the most common infectious disease of cattle. This has led to high economic losses in the cattle farming industry. BoHV-1 can potentially be transmitted via semen during natural or artificial insemination (AI). Therefore, testing methods for the early diagnosis of BoHV-1 infection are urgently needed for international trade of ruminant semen. In this study, we developed a novel droplet digital PCR (ddPCR) assay for the detection of BoHV-1 DNA in semen samples.
RESULTS
The ddPCR results showed that the detection limit was 4.45 copies per reaction with high reproducibility. The established method was highly specific for BoHV-1 and did not show cross-reactivity with specify the organisms (BTV, BVDV, Brucella, M . bovis). The results of clinical sample testing showed that the positivity rate of ddPCR (87.8%) was higher than that of qPCR (84.1%).
CONCLUSIONS
The ddPCR assay showed good accuracy for mixed samples and could be a new added diagnostic tool for detecting BoHV-1.
背景
由牛疱疹病毒 1 型（BoHV-1）引起的传染性牛鼻气管炎（IBR）是牛的最重要传染病之一。这是牛最常见的传染病之一。这导致了养牛业的巨大经济损失。BoHV-1 可以通过自然或人工授精（AI）期间的精液传播。因此，迫切需要用于检测 BoHV-1 感染的早期诊断的检测方法，以便进行反刍动物精液的国际贸易。在这项研究中，我们开发了一种新的液滴数字 PCR（ddPCR）检测方法，用于检测精液样本中的 BoHV-1 DNA。
结果
ddPCR 结果表明，检测限为每个反应 4.45 拷贝，具有很高的重现性。该方法高度特异性针对 BoHV-1，与特定生物体（BTV、BVDV、布鲁氏菌、牛分枝杆菌）没有交叉反应。临床样本检测结果表明，ddPCR 的阳性率（87.8%）高于 qPCR（84.1%）。
结论
ddPCR 检测方法对混合样本具有良好的准确性，可能成为检测 BoHV-1 的新的附加诊断工具。
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